Sequence of a yeast DNA fragment containing a chromosomal replicator and the TRP1 gene

Tschumper, Gary; Carbon, John

doi:10.1016/0378-1119(80)90133-x

Cited by 510 publications

(200 citation statements)

References 21 publications

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“…However, the nucleotide sequence of the cloned fragment reveals a sequence that is closely related to the consensus core sequence, -4'TTTATPuTT T, found within identified ARS elements (4,18,44,47). The sequence -ATTTGTATTT-l , complementary to nucleotides 1795 to 1805 in the 3'-flanking region of the CDC7 gene, differs by only one nucleotide from the ARS core consensus.…”

Section: Resultsmentioning

confidence: 94%

Molecular characterization of cell cycle gene CDC7 from Saccharomyces cerevisiae.

Patterson¹,

Sclafani²,

Fangman³

et al. 1986

Mol. Cell. Biol.

119

101

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The product of the CDC7 gene of Saccharomyces cerevisiae appears to have multiple roles in cellular physiology. It is required for the initiation of mitotic DNA synthesis. While it is not required for the initiation of meiotic DNA replication, it is necessary for genetic recombination during meiosis and for the formation of ascospores. It has also been implicated in an error-prone DNA repair pathway. Plasmids Use of the budding yeast Saccharomyces cerevisiae as a model for studies on the eucaryotic cell cycle relies heavily on temperature-sensitive mutations in cell division cycle (CDC) genes (32, 35). Characterization of these mutants has led to the formulation of a model in which progression through the cell cycle is determined by a set of interrelated pathways, each organized as a dependent sequence of events requiring the action of specific gene products. One such pathway, operating late in the Gi phase of the cell cycle, requires the function of the CDC7 gene product (12). Cells carrying a thermosensitive lesion in the CDC7 gene arrest at the restrictive temperature as budded cells with separated spindle-pole bodies but without an elongated spindle apparatus and without initiating DNA synthesis (5, 12). Upon return to permissive conditions cdc7 cells are able to enter the S phase and subsequently complete a round of DNA synthesis without further protein synthesis (14).In contrast to the requirement for CDC7 function to initiate mitotic DNA synthesis, premeiotic DNA replication occurs normally in cdc7 homozygous diploids under the restrictive condition (43). However, these diploids fail to form a synaptonemal complex, to show commitment to genetic recombination, or to form ascospores (40). Thus, although cdc7 strains are defective in both mitotic and meiotic cell cycles, the lesion appears to affect each pathway in a quite distinct manner. In addition to having roles in the mitotic and meiotic pathways, the CDC7 gene product has been implicated in DNA repair as a member of the RAD6 epistasis group, since strains carrying a cdc7 mutation show almost no mutagenic repair in response to a variety of damaging agents (31).To elucidate the role of the CDC7 gene product in the various cellular functions in which it is implicated and to determine whether differential expression of the CDC7 gene is involved with its cell cycle functions, we and others (24) have begun a molecular analysis of the CDC7 gene. In this paper we describe the cloning of the CDC7 gene, the characterization of its transcriptional product, the nucleotide sequence of the gene, and the regions of homology between the predicted protein products of the CDC7 and CDC28 genes. MATERIALS AND METHODSStrains and media. Escherichia coli HB101 (F-thi leu pro hsdR hsdM recA end!) and HW87 [F-A(araD139-leu) lacX74 galK hsdR rpsL srb recA] were used as hosts for the routine maintenance and propagation of plasmids. Bacterial cultures were grown in L broth or supplemented M9 medium (26); when necessary, ampicillin was added to media to a final concentration of...

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Section: Resultsmentioning

confidence: 94%

Molecular characterization of cell cycle gene CDC7 from Saccharomyces cerevisiae.

Patterson¹,

Sclafani²,

Fangman³

et al. 1986

Mol. Cell. Biol.

119

101

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“…Plasmids in Saccharomyces cerevisiae which contain an autonomous replicating sequence (ARS) and a centromeric DNA element (CEN) are replicated and segregated in a manner similar to chromosomes (18,(20)(21)(22).…”

Section: Resultsmentioning

confidence: 99%

“…Figure 1 shows projections for the synthetic oligonucleotides (A5N5) 20 (A8N2) 20 (7), kinetoplast DNA (10), human ARS2 (17), and yeast centromeric element CEN3 (18). The programs also calculate the ratio of the contour length of the axis of a given molecule to the shortest distance between its ends.…”

Section: Methodsmentioning

confidence: 99%

Computer modelling of DNA structures involved in chromosome maintenance

Eckdahl

Anderson

1987

Nucl Acids Res

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Sequence-dependent DNA bending of synthetic and natural molecules was studied by computer analysis.Modelling of synthetic oligonucleotides and of 107 kb of natural sequences gave results which closely resembled published electrophoretic data, demonstrating the powerful predictive capacity of the procedure.The analysis was extended to the study of DNA structures involved in chromosome maintenance.Centromeric DNAs from yeast were found to have sequences in their functional elements which cause them to be unusually straight. Autonomous replicating sequences were found to have two structural domains, one consisting of unusually straight sequences surrounding the consensus and the other of bending elements in flanking DNA. In addition to a structural homology, centromeric and autonomous replicating sequences share common sequence elements.These observations show that computer modelling of natural sequences is a viable approach to the study of the biological implications of alternative DNA structures.

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“…Yeast replication plasmids, YRp7 and pJJ283, a single copy plasmid, pRS314, and multicopy plasmids, pYO324 and pSQ326, were described previously (Tschumper and Carbon, 1980;Sikorski and Hieter, 1989;Jones and Prakash, 1990;Ohya et al, 1991;Qadota et al, 1992). pSHF9-4 [2, SEC12-MFal URA3 I was constructed as described (Nishikawa and Nakano, 1993).…”

Section: Plasmids and Dna Manipulationsmentioning

confidence: 99%

Membrane protein retrieval from the Golgi apparatus to the endoplasmic reticulum (ER): characterization of the RER1 gene product as a component involved in ER localization of Sec12p.

Sato

Nishikawa

Nakano

1995

MBoC

101

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Yeast Sec12p, a type II transmembrane glycoprotein, is required for formation of transport vesicles from the endoplasmic reticulum (ER). Biochemical and morphological analyses have suggested that Sec12p is localized to the ER by two mechanisms: static retention in the ER and dynamic retrieval from the early region of the Golgi apparatus. The rer1 mutant we isolated in a previous study mislocalizes the authentic Sec12p to the later compartments of the Golgi. To understand the role of RER1 on Sec12p localization, we cloned the gene and determined its reading frame. RER1 encodes a hydrophobic protein of 188 amino acid residues containing four putative membrane spanning domains. The rer1 null mutant is viable. Even in the rer1 disrupted cells, immunofluorescence of Sec12p stains the ER, implying that the retention system is still operating in the mutant. To determine the subcellular localization of Rer1p, an epitope derived from the influenza hemagglutinin was added to the C-terminus of Rer1p and the cells expressing this tagged but functional protein were observed by immunofluorescence microscopy. The anti-HA monoclonal antibody stains the cells in a punctate pattern that is typical for Golgi proteins and clearly distinct from the ER staining. This punctate staining was in fact exaggerated in the sec7 mutant that accumulates the Golgi membranes at the restrictive temperature. Furthermore, double staining of Rer1p and Ypt1p, a GTPase that is known to reside in the Golgi apparatus, showed good colocalization. Subcellular fractionation experiments indicated that the fractionation pattern of Rer1p was similar to that of an early Golgi protein, Och1p. From these results, we suggest that Rer1p functions in the Golgi membrane to return Sec12p that has escaped from the static retention system of the ER.

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Sequence of a yeast DNA fragment containing a chromosomal replicator and the TRP1 gene

Cited by 510 publications

References 21 publications

Molecular characterization of cell cycle gene CDC7 from Saccharomyces cerevisiae.

Molecular characterization of cell cycle gene CDC7 from Saccharomyces cerevisiae.

Computer modelling of DNA structures involved in chromosome maintenance

Membrane protein retrieval from the Golgi apparatus to the endoplasmic reticulum (ER): characterization of the RER1 gene product as a component involved in ER localization of Sec12p.

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